Anti-MTAP antibody [EPR22570-76] (ab254265)

Anti-MTAP antibody [EPR22570-76] (ab254265) is a rabbit monoclonal antibody that is used to detect MTAP in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat samples.

- Specificity confirmed with MTAP knockout cell line validation

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Related conjugates and formulations

ab256356
Carrier free
Anti-MTAP antibody [EPR22570-76] - BSA and Azide free
ab306398
Conjugated
APC Anti-MTAP antibody [EPR22570-76]
ab306399
Conjugated
HRP Anti-MTAP antibody [EPR22570-76]
ab309821
Conjugated
Alexa Fluor® 488 Anti-MTAP antibody [EPR22570-76]
ab310190
Conjugated
Alexa Fluor® 647 Anti-MTAP antibody [EPR22570-76]
ab310631
Conjugated
Alexa Fluor® 594 Anti-MTAP antibody [EPR22570-76]
ab312645
Conjugated
Alexa Fluor® 568 Anti-MTAP antibody [EPR22570-76]
ab306397
Conjugated
PE Anti-MTAP antibody [EPR22570-76]
ab312158
Conjugated
Alexa Fluor® 555 Anti-MTAP antibody [EPR22570-76]
ab321502
Conjugated
Alexa Fluor® 750 Anti-MTAP antibody [EPR22570-76]

Images

Western blot - Anti-MTAP antibody [EPR22570-76] (AB254265), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Not recommended	Tested	Tested	Tested	Tested
Mouse	Not recommended	Tested	Expected	Expected	Tested
Rat	Not recommended	Tested	Expected	Expected	Tested

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/400	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

See full target information MTAP

Function

Catalyzes the reversible phosphorylation of S-methyl-5'-thioadenosine (MTA) to adenine and 5-methylthioribose-1-phosphate. Involved in the breakdown of MTA, a major by-product of polyamine biosynthesis. Responsible for the first step in the methionine salvage pathway after MTA has been generated from S-adenosylmethionine. Has broad substrate specificity with 6-aminopurine nucleosides as preferred substrates.

Alternative names

MSAP, MTAP, S-methyl-5'-thioadenosine phosphorylase, 5'-methylthioadenosine phosphorylase, MTA phosphorylase, MTAPase

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR22570-76
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What is this antibody validated in?

Anti-MTAP antibody [EPR22570-76] (ab254265) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human, Mouse, Rat samples.

What is the molecular weight of MTAP?

Anti-MTAP [EPR22570-76] (ab254265) specifically detects a band for MTAP (UniProt: Q13126) at a molecular weight of 31kDa.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Specificity confirmed

The specificity of Anti-MTAP antibody [EPR22570-76] (ab254265) has been confirmed by Western blot testing in MTAP Knockout HeLa cell line, ab265272.

Other related products

We have a range of other formats of antibody clone [EPR22570-76] also available for your convenience:
ab254265, Carrier free - ab256356, PE - ab306397, APC - ab306398, HRP - ab306399, Alkaline Phosphatase - ab308905, Alexa Fluor® 488 - ab309821, Alexa Fluor® 647 - ab310190, Alexa Fluor® 594 - ab310631, Alexa Fluor® 555 - ab312158, Alexa Fluor® 568 - ab312645, Alexa Fluor® 750 - ab321502

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

MTAP also known as methylthioadenosine phosphorylase is an essential enzyme that breaks down 5'-methylthioadenosine (MTA) a byproduct of polyamine synthesis. MTAP has a molecular weight of about 31 kDa. The enzyme converts MTA into adenine and 5-methylthioribose-1-phosphate which re-enter the methionine and adenine salvage pathways. MTAP is widely expressed in most tissues but its activity is especially high in the liver and kidney. Alternative names for MTAP include 2G4 and MTAP-A.

Biological function summary

Methylthioadenosine phosphorylase plays a significant role in the salvage pathways for methionine and adenine critical for cellular growth and proliferation. MTAP operates as a part of a complex metabolic network involved in polyamine metabolism. In addition to its metabolic functions MTAP contributes to the regulation of the immune response and cell cycle. MTAP immunohistochemistry is often used to study its expression patterns in various tissues.

Pathways

Methylthioadenosine phosphorylase participates importantly in the polyamine biosynthesis and methionine salvage pathways. These pathways are integral for maintaining cellular homeostasis and nucleotide pools. MTAP works closely with proteins such as methionine adenosyltransferase (MAT) and adenosylmethionine decarboxylase (AMD). In concert they facilitate the regeneration of methionine highlighting MTAP's role in cellular adaptation to metabolic demands.

Associated diseases and disorders

Methylthioadenosine phosphorylase deficiency or deletion is linked to certain cancers such as gliomas and lymphomas. Loss of MTAP function is often associated with co-deletion of the tumor suppressor protein p16INK4a observed in various malignancies. This deletion can lead to an accumulation of MTA creating a toxic environment that promotes cancer cell proliferation. MTAP and its interaction with proteins like p16INK4a highlight its relevance as a potential target for therapeutic intervention in cancer treatment programs.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265)
Lanes 1-4: Merged signal (red and green). Green - ab254265 observed at 32 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
ab254265 Anti-MTAP antibody [EPR22570-76] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human MTAP knockout HeLa cell line ab265272 (knockout cell lysate Human MTAP knockout HeLa cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. ab254265 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: MTAP knockout HeLa cell lysate at 20 µg
Lane 3: HT-29 cell lysate at 20 µg
Lane 4: A549 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 32 kDa
Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265)
Lanes 1-4: Merged signal (red and green). Green - ab254265 observed at 32 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
ab254265 Anti-MTAP antibody [EPR22570-76] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human MTAP knockout HeLa cell line ab265272 (knockout cell lysate Human MTAP knockout HeLa cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. ab254265 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: MTAP knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human MTAP knockout HeLa cell line (Human MTAP knockout HeLa cell line ab265272)
Lane 3: HT-29 cell lysate at 20 µg
Lane 4: A549 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 32 kDa
Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265)
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times:
Lanes 1-4: 15 seconds; Lane 5: 10 seconds; Lane 6: 3.25 seconds.
The expression molecular weight observed is consistent with what has been described in the literature (PMID:15492751).
All lanes: Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265) at 1/1000 dilution
Lane 1: Mouse skin lysate at 20 µg
Lane 2: Mouse liver lysate at 20 µg
Lane 3: Rat skin lysate at 20 µg
Lane 4: Rat liver lysate at 20 µg
Lane 5: RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 20 µg
Lane 6: NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 31 kDa
Observed band size: 32 kDa
Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265)
anes 1- 4: Merged signal (red and green). Green - Anti-MTAP antibody [EPR6893] ab126770 observed at 32 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
Anti-MTAP antibody [EPR6893] ab126770 Anti-MTAP antibody [EPR6893] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human MTAP knockout HeLa cell line ab265272 (knockout cell lysate Human MTAP knockout HeLa cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. Anti-MTAP antibody [EPR6893] ab126770 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: MTAP knockout HeLa cell lysate at 20 µg
Lane 3: HT-29 cell lysate at 20 µg
Lane 4: A549 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 32 kDa
Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265)
Blocking and dilution buffer: 5% NFDM/TBST.
Negative control: A549 (PMID: 17548352).
The expression molecular weight observed is consistent with what has been described in the literature (PMID:15492751).
Exposure times.
Lanes 1-2: 10 seconds; Lane3: 5.5 seconds; Lanes 4-8: 3.25 seconds.
All lanes: Western blot - Anti-MTAP antibody [EPR22570-76] (ab254265) at 1/1000 dilution
Lane 1: Human liver lysate at 20 µg
Lane 2: Human placenta lysate at 20 µg
Lane 3: HT-29 (human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 4: HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 5: HeLa (human epithelial cell line from cervix adenocarcinoma0 whole cell lysate at 20 µg
Lane 6: Raji 9human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Lane 7: Caco-2 (human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 8: A549 9human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg
Secondary
Lanes 1 - 2: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution
Lanes 3 - 8: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 31 kDa
Observed band size: 32 kDa
Flow Cytometry (Intracellular) - Anti-MTAP antibody [EPR22570-76] (ab254265)
Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized A549 (human lung carcinoma cell line) (Left) anf H-29 (human colorectal adenocarcinoma cell line) (Right) cells labeling MTAP with ab254265 at 1/400 dilution (red) compared with Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).
Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), at 1/2000 dilution was used as the secondary antibody.
Negative control: A549 (PMID: 8971171.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR22570-76] (ab254265)
Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling MTAP with ab254265 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in rat liver (PMID:15492751) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR22570-76] (ab254265)
Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling MTAP with ab254265 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in mouse liver (PMID:15492751) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR22570-76] (ab254265)
Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling MTAP with ab254265 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in stroma cells and no staining in tumor cells of human bladder cancer (PMID:27270441) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR22570-76] (ab254265)
Immunohistochemical analysis of paraffin-embedded human breast tissue labeling MTAP with ab254265 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining in human breast (PMID:26751376, 18712977) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Immunocytochemistry/ Immunofluorescence - Anti-MTAP antibody [EPR22570-76] (ab254265)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (human lung carcinoma cell line) and HT-29 (human colorectal adenocarcinoma cell line) cells labeling MTAP with ab254265 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and nuclear staining in HT-29 cell line. The nuclear counter stain is DAPI (blue) Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 dilution (red).
SEcondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
Negative control: A549（PMID: 8971171.